Seasonal fire effects on the diversity patterns,spatial distribution and community structure of forbs in the Northern Mixed Prairie,USA

The effects of fire season on forb diversity patterns, density, and composition were determined for a northern Mixed Prairie site, USA. Repeated spring burns (dormant season), summer burns, fall burns (dormant season), and unburned treatments were compared over a 3-yr period characterized by wet and dry moisture conditions. Alpha and beta diversity were highest on unburned and summer burn treatments, while landscape mosaic diversity was highest on fall burns. Forb density was highest on fall and spring burn sites. Nine forb species comprised 82% of total densities and were significantly affected by fire season and year to year variations in moisture. Forb composition for unburned and spring burn treatments was similar, but both treatments were different from the summer burn and fall burn treatments which were similar to each other. Fire alone did not appear to be an intense enough disturbance to initiate drastic changes in the forb component of vegetation patches. Specific fire seasons did appear to either mask or enhance forb structure arising from other disturbance(s). Fire season also affected the scales of forb organization in the landscape. Contrasting spatial characteristics of the forb component of prairie plant communities may provide a diagnostic technique for exposing the interaction of disturbances at different temporal and spatial scales.     

Antiestrogenic and anticancer activities of peptides derived from the active site of alpha‐fetoprotein

Cyclo[EKTOVNOGN] (AFPep), a cyclic 9‐amino acid peptide derived from the active site of alpha‐fetoprotein, has been shown to prevent carcinogen‐induced mammary cancer in rats and inhibit the growth of ER⁺ human breast cancer xenografts in mice. Recently, studies using replica exchange molecular dynamics predicted that the TOVN region of AFPep might form a dynamically stable putative Type I beta‐turn, and thus be biologically active without additional amino acids. The studies presented in this paper were performed to determine whether TOVN and other small analogs of AFPep would inhibit estrogen‐stimulated cancer growth and exhibit a broad effective‐dose range. These peptides contained nine or fewer amino acids, and were designed to bracket or include the putative pharmacophoric region (TOVN) of AFPep. Biological activities of these peptides were evaluated using an immature mouse uterine growth inhibition assay, a T47D breast cancer cell proliferation assay, and an MCF‐7 breast cancer xenograft assay. TOVN had very weak antiestrogenic activity in comparison to AFPep's activity, whereas TOVNO had antiestrogenic and anticancer activities similar to AFPep. OVNO, which does not form a putative Type I beta‐turn, had virtually no antiestrogenic and anticancer activities. A putative proteolytic cleavage product of AFPep, TOVNOGNEK, significantly inhibited E₂‐stimulated growth in vivo and in vitro over a wider dose range than AFPep or TOVNO. We conclude that TOVNO has anticancer potential, that TOVNOGNEK is as effective as AFPep in suppressing growth of human breast cancer cells, and that it does so over a broader effective‐dose range. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd.     

Fine‐scale heterogeneity in beetle assemblages under co‐occurring Eucalyptus in the same subgenus

Aim Insect biodiversity is often positively associated with habitat heterogeneity. However, this relationship depends on spatial scale, with most studies focused on differences between habitats at large scales with a variety of forest tree species. We examined fine‐scale heterogeneity in ground‐dwelling beetle assemblages under co‐occurring trees in the same subgenus: Eucalyptus melliodora A. Cunn. ex Schauer and E. blakelyi Maiden (Myrtaceae). Location Critically endangered grassy woodland near Canberra, south‐eastern Australia. Methods We used pitfall traps and Tullgren funnels to sample ground‐dwelling beetles from the litter environment under 47 trees, and examined differences in diversity and composition at spatial scales ranging from 100 to 1000 m. Results Beetle assemblages under the two tree species had distinctive differences in diversity and composition. We found that E. melliodora supported a higher richness and abundance of beetles, but had higher compositional similarity among samples. In contrast, E. blakelyi had a lower abundance and species richness of beetles, but more variability in species composition among samples. Main conclusions Our study shows that heterogeneity in litter habitat under co‐occurring and closely related eucalypt species can influence beetle assemblages at spatial scales of just hundreds of metres. The differential contribution to fine‐scale alpha and beta diversity by each eucalypt can be exploited for conservation purposes by ensuring an appropriate mix of the two species in the temperate woodlands where they co‐occur. This would help not only to maximize biodiversity at landscape scales, but also to maintain heterogeneity in species richness, trophic function and biomass at fine spatial scales.     

223Ra-dichloride spectrometric characterization: Searching for the presence of long-lived isotopes with radiological protection implications

²²³Ra-dichloride was approved with the commercial name of Xofigo in 2014 for treatment of metastatic castration-resistant prostate cancer.²²³Ra is obtained by neutron irradiation of ²²⁶Ra yielding ²²⁷Ac, which decays to ²²⁷Th and ²²³Fr, both decaying to ²²³Ra. Since ²²³Ra is predominantly (95.3%) an alpha emitter with a 11.42 days long half-life, the radiopharmaceutical, its remnants, the patient, and waste material can be managed and disposed with low radiation protection requirements.²²⁷Ac is a long-lived (T_1/2 = 21.77 years) beta emitter that demands strong radiation protection measures. In particular waste disposal has to follow the International Atomic Energy Agency (IAEA) and European Commission (EC) regulations. Since ²²⁷Ac is involved in the production of ²²³Ra, an impurity analysis of each batch is required after production. Due to time restrictions, the manufacturer’s detection limit (<0.001%) exceeds the one required to assure that ²²⁷Ac concentrations are below direct disposal levels. To improve the detection limit, long-term accurate spectroscopy is required.Alpha and gamma spectroscopy measurements were carried out at the Complutense University Nuclear Physics Laboratory. After twelve months follow up of a sample, ²²⁷Ac concentration was found to be smaller than 10⁻⁹. This allows for direct waste disposal and no additional radiation protection restrictions than those required for ²²³Ra. The presence of contamination by other radioisotopes was also ruled out by this experiment. Specifically ²²⁶Ra, involved in ²²³Ra production as the original parent and with a very long-lived (T_1/2 = 1577 years) alpha emitter, was also below the experimental detection limit.     

Characterization of the C584R variant in the mtDNA depletion syndrome gene FBXL4, reveals a novel role for FBXL4 as a regulator of mitochondrial fusion

Mutations in FBXL4 (F-Box and Leucine rich repeat protein 4), a nuclear-encoded mitochondrial protein with an unknown function, cause mitochondrial DNA depletion syndrome. We report two siblings, from consanguineous parents, harbouring a previously uncharacterized homozygous variant in FBXL4 (c.1750 T > C; p.Cys584Arg). Both patients presented with encephalomyopathy, lactic acidosis and cardiac hypertrophy, which are reported features of FBXL4 impairment. Remarkably, dichloroacetate (DCA) administration to the younger sibling improved metabolic acidosis and reversed cardiac hypertrophy. Characterization of FBXL4 patient fibroblasts revealed severe bioenergetic defects, mtDNA depletion, fragmentation of mitochondrial networks, and abnormalities in mtDNA nucleoids. These phenotypes, observed with other pathogenic FBXL4 variants, confirm the pathogenicity of the p.Cys584Arg variant. Although treating FBXL4 fibroblasts with DCA improved extracellular acidification, in line with reduced lactate levels in patients, DCA treatment did not improve any of the other mitochondrial functions. Nonetheless, we highlight DCA as a potentially effective drug for the management of elevated lactate and cardiomyopathy in patients with pathogenic FBXL4 variants. Finally, as the exact mechanism through which FBXL4 mutations lead to mtDNA depletion was unknown, we tested the hypothesis that FBXL4 promotes mitochondrial fusion. Using a photo-activatable GFP fusion assay, we found reduced mitochondrial fusion rates in cells harbouring a pathogenic FBXL4 variant. Meanwhile, overexpression of wildtype FBXL4, but not the p.Cys584Arg variant, promoted mitochondrial hyperfusion. Thus, we have uncovered a novel function for FBXL4 in promoting mitochondrial fusion, providing important mechanistic insights into the pathogenic mechanism underlying FBXL4 dysfunction.     

NMR assignment of S836: a de novo protein from a designed superfamily

Libraries of de novo proteins provide an opportunity to explore the structural potential of biological macromolecules that have not been biased by billions of years of evolutionary selection. Characterization of individual members of such libraries provides insight into the diversity of structure and dynamics accessible to nascent protein superfamilies in the absence of evolutionary optimization. Here we report the backbone and side chain chemical shifts of protein S836 from a superfamily of designed 4-helix bundles.     

Heavy chains of inter alpha inhibitor (IαI) inhibit the human complement system at early stages of the cascade

Inter alpha inhibitor (IαI) is an abundant serum protein consisting of three polypeptides: two heavy chains (HC1 and HC2) and bikunin, a broad-specificity Kunitz-type proteinase inhibitor. The complex is covalently held together by chondroitin sulfate but during inflammation IαI may interact with TNF-stimulated gene 6 protein (TSG-6), which supports transesterification of heavy chains to hyaluronan. Recently, IαI was shown to inhibit mouse complement in vivo and to protect from complement-mediated lung injury but the mechanism of such activity was not elucidated. Using human serum depleted from IαI, we found that IαI is not an essential human complement inhibitor as was reported for mice and that such serum has unaltered hemolytic activity. However, purified human IαI inhibited classical, lectin and alternative complement pathways in vitro when added in excess to human serum. The inhibitory activity was dependent on heavy chains but not bikunin and detected at the level of initiating molecules (MBL, properdin) in the lectin/alternative pathways or C4b in the classical pathway. Furthermore, IαI affected formation and assembly of the C1 complex and prevented assembly of the classical pathway C3-convertase. Presence and putative interactions with TSG-6 did not affect the ability of IαI to inhibit complement thus implicating IαI as a potentially important complement inhibitor once enriched onto hyaluronan moieties in the course of local inflammatory processes. In support of this, we found a correlation between IαI/HC-containing proteins and hemolytic activity of synovial fluid from patients suffering from rheumatoid arthritis.     

Purification and characterization of Hb 98-114: A novel hemoglobin-derived antimicrobial peptide from the midgut of Rhipicephalus (Boophilus) microplus

The antimicrobial activity of hemoglobin fragments (hemocidins) has been reported in a variety of models. The cattle tick Rhipicephalus (Boophilus) microplus is a blood sucking arthropod from where the first in vivo-generated hemocidin was characterized (Hb 33-61). In the present work we identified a novel antimicrobial peptide from the midgut of fully engorged R. (B.) microplus females, which comprises the amino acids 98-114 of the alpha subunit of bovine hemoglobin, and was designated Hb 98-114. This peptide was active against several yeast and filamentous fungi, although no activity was detected against bacteria up to 50μM of the synthetic peptide. Hb 98-114 was capable of permeabilizing Candida albicans cell membrane and had a fungicidal effect against this yeast. Circular dichroism (CD) and nuclear magnetic resonance (NMR) experiments showed that Hb 98-114 has a random conformation in aqueous solution but switches to an alpha-helical conformation in the presence of sodium dodecyl sulfate (SDS). This alpha helix adopts an amphipathic structure which may be the mechanism of cell membrane permeabilization. Importantly, Hb 98-114 may play an important role in defending the tick midgut against fungal pathogens and is the first hemocidin with specific antifungal activity to be characterized.     

Arctiid moth ensembles along a successional gradient in the Ecuadorian montane rain forest zone: how different are subfamilies and tribes?

Aim We examined changes in the species diversity and faunal composition of arctiid moths along a successional gradient at a fine spatial scale in one of the world's hot spots for moths, the Andean montane rain forest zone. We specifically aimed to discover whether moth groups with divergent life histories respond differentially to forest recovery. Location Southern Ecuador (province Zamora‐Chinchipe) along a gradient from early successional stages to mature forest understorey at elevations of 1800–2005 m a.s.l. Methods Moths were sampled with weak light traps at 21 sites representing three habitat categories (early and late succession, mature forest understorey), and were analysed at species level. Relative proportions were calculated from species numbers as well as from specimen numbers. Fisher's α was used as a measure of local diversity, and for ordination analyses non‐metric multidimensional scaling (NMDS) was carried out. Results Proportions of higher arctiid taxa changed distinctly along the successional gradient. Ctenuchini (wasp moths) contributed more strongly to ensembles in natural forest, whereas Lithosiinae (lichen moths) decreased numerically with forest recovery. Arctiid species diversity (measured as Fisher's α) was high in all habitats sampled. The three larger subordinated taxa contributed differentially to richness: Phaegopterini (tiger moths) were always the most diverse clade, followed by Ctenuchini and Lithosiinae. Local species diversity was higher in successional habitats than in forest understorey, and this was most pronounced for the Phaegopterini. Dominance of a few common species was higher, and the proportion of species represented as singletons was lower, than reported for many other tropical arthropod communities. NMDS revealed a significant segregation between ensembles from successional sites and from forest understorey for all larger subordinated taxa (Phaegopterini, Ctenuchini, Lithosiinae). Abandoned pastures held an impoverished, distinct fauna. Faunal segregation was more pronounced for rare species. Ordination axes reflected primarily the degree of habitat disturbance (openness of vegetation, distance of sites from mature forest) and, to a lesser extent, altitude, but not distance between sampling sites. Main conclusions Despite the geographical proximity of the 21 sites and the pronounced dispersal abilities of adult arctiid moths, local ecological processes were strong enough to allow differentiation between ensembles from mature forest and disturbed sites, even at the level of subfamilies and tribes. Differences in morphology and life‐history characteristics of higher arctiid taxa were reflected in their differential representation (proportions of species and individuals) at the sites, whereas patterns of alpha and beta diversity were concordant. However, concordance was too low to allow for reliable extrapolation, in terms of biodiversity indication, from one tribe or subfamily to the entire family Arctiidae. Phaegopterini (comprising more putative generalist feeders during the larval stages) benefited from habitat disturbance, whereas Ctenuchini (with host‐specialist larvae) were more strongly affiliated with forest habitats.